Calr条件敲除小鼠模型的构建
Construction of Calr conditional knockout mice model
摘要
目的：构建条件性敲除Calr基因的小鼠模型。Calr是一种定位于内质网中的钙结合蛋白，被认为是内质网中的关键蛋白伴侣，此模型的建立可以为研究内质网中的蛋白质稳态提供重要实验基础。方法：利用CRISPR/Cas9 基因编辑技术构建Calr 4-7 exon靶向敲除小鼠，再与Mx1-Cre小鼠繁殖得到所需要小鼠。在小鼠年龄6周到8周时，对小鼠poly I:C腹腔注射诱导cre表达，注射完两周之后对小鼠进行PCR鉴定与Western blot蛋白印迹实验，从DNA水平与蛋白质水平上证明实现了Calr基因的成功敲除。结论：成功构建可在体内造血系统里条件性敲除Calr的小鼠模型，为后续基于蛋白质稳态调控的角度以提高造血干细胞体外扩增应用提供了新思路。
Abstract
Objective: To construct a mouse model with conditional knockout of the Calr gene. Calr is a calcium-binding protein located in the endoplasmic reticulum and is considered a key protein chaperone in the endoplasmic reticulum. Establishing this model can provide an important experimental basis for studying protein homeostasis in the endoplasmic reticulum. Methods: Using CRISPR/Cas9 gene editing technology, mice with targeted knockout of Calr exons 4-7 were constructed, and then bred with Mx1-Cre mice to obtain the required mice. At 6 to 8 weeks of age, the mice were intraperitoneally injected with poly I:C to induce cre expression. Two weeks after injection, PCR identification and Western blot protein analysis were performed on the mice to demonstrate successful knockout of the Calr gene at both the DNA and protein levels. Conclusion: A mouse model with conditional knockout of Calr in the hematopoietic system in vivo was successfully constructed, providing a new approach for subsequent studies on protein homeostasis regulation to enhance the in vitro expansion application of hematopoietic stem cells.关键词
分子医学/造血干细胞/内质网蛋白质稳态/CalrKey words
Molecular medicine/hematopoietic stem cells/Endoplasmic Reticulum Protein Homeostasis/Calr引用本文复制引用
刘苏萱,刘禄.Calr条件敲除小鼠模型的构建[EB/OL].(2026-03-27)[2026-03-29].http://www.paper.edu.cn/releasepaper/content/202603-263.学科分类
基础医学/生物科学理论、生物科学方法/生物科学研究方法、生物科学研究技术
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